UGT1A10基因SNP rs10187694对吗替麦考酚酯代谢的影响  

Effect of SNP rs10187694 of UGT1A10 gene on metabolism of Mycophenolate mofetil

作　　者：肖超[1] 孙红成[1] 邓贵龙[1] 宋国贺 王宇鹏[1] 陈健[1] 孙星[1] 钟林[1] 彭志海[1] 王晓亮[1] 

机构地区：[1]上海交通大学附属第一人民医院普外科,上海200080

出　　处：《中国现代医学杂志》2017年第12期9-14,共6页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:81270557;81110108010)

摘　　要：目的体外研究尿苷二磷酸葡萄糖醛酸转移酶(UGT1A10)基因SNP rs10187694多态位点G/A变异对吗替麦考酚酯(MMF)代谢的影响,明确该位点变异是否与MMF个体代谢差异有关。方法采用基因重组、定点突变技术,构建UGT1A10基因SNP rs10187694位点含有不同等位基因的重组过表达载体,POLO 3000转染法将重组过表达载体转染入HEK293细胞,采用液相色谱-质谱联用仪(LC/MS/MS)系统,检测霉酚酸(MPA)代谢产物7-O-葡醛酸苷(MPAG)的生成量,以评估转染不同等位基因HEK293细胞中酶的活性。结果成功构建pIRES2-EGFP-prom(G)和pIRES2-EGFP-prom(A)重组过表达载体,并转染HEK293细胞。LC/MS/MS系统检测结果显示,携带A等位基因的突变型UGT1A10代谢MMF产生MPAG的能力降低,其24 h MPAG的生成量为(226.00±14.57)nmol/L,而野生型的生成量为(269.00±14.07)nmol/L,UGT1A10突变型MPAG的生成量是野生型的84.00%,两者比较,差异有统计学意义(P<0.05)。结论 UGT1A10基因SNP rs10187694位点G/A突变可影响MMF代谢产物MPAG的生成,可能是造成不同个体MMF代谢差异的重要原因之一。Objective To study the effect of G/A mutation in SNP rs10187694 site of uridine diphosphate glucuronosyl transferase （UGTIAIO） on Myeophenolate mofetil metabolism. Methods The recombinant overex- pression vectors of UGT1AIO gene were constructed by gene recombination technique and the strategy of site directed mutagenesis. Then, the recombinant plasmid was transfected into HEK293 cells by POLO3000 method. By detecting the yields of myeophenolic acid （MPA） metabolite 7-o-glucuronide （MPAG）, the activity of enzyme in the HEK293 cells transfected with different alleles were evaluated. Results The overexperession vectors plRES2-EGFP-prom （G） and plRES2-EGFP-prom （A） were successfully constructed and transfeeted into HEK293 cells. The results analyzed by LC/MS/MS showed that the cells containing mutant type of UGT1A10 gene with A allele produced less MPAG than those with G allele, the concentration of MPAG produced by mutant type in 24 hours was （226.00± 14.57） nmol/L, however, the concentration in wild type was （269.00 ± 14.07） nmol/L. The generation amount of MPAG in mutant type was 84.00% of that in wild type in 24 hours （P 〈 0.05）. Conclusions UGT1AIO gene SNP rs10187694 can significantly affect the production of MPAG,which may be one of the reasons causing the difference in Mycophenolate mofetil metabolism.

关 键 词：尿苷二磷酸葡萄糖醛酸转移酶 霉酚酸酯 药物代谢 

分 类 号：R969.1[医药卫生—药理学]